Roles of Macropinocytosis in HIV-1 infection of CD4+ T Cells

巨胞饮作用在 HIV-1 感染 CD4 T 细胞中的作用

• 批准号: 10412160
• 负责人: PHILIP D KING
• 金额: $ 65.25万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-23 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10412160
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Binding; Biology; CCR5 gene; CD4 Positive T Lymphocytes; CXCR4 gene; Capsid; Cell Line; Cell Nucleus; Cell fusion; Cell membrane; Cell physiology; Cell surface; Cells; Clathrin; Complex; Conflict (Psychology); Cytoplasm; Data; Endocytosis; Environment; Event; Genes; Genetic; Goals; HIV; HIV Envelope Protein gp120; HIV Infections; HIV-1; Human; Immunology; Infection; Knowledge; Laboratories; Location; Mediating; Metabolism; Microscopic; Molecular; Molecular Conformation; N-terminal; Nature; Outcome; Pathway interactions; Peptides; Pharmacology; Play; Primary Infection; Process; Property; Reporting; Research; Role; Signal Transduction; Surface; T-Lymphocyte; Testing; Viral; Virion; Virus; Virus Diseases; cell type; env Glycoproteins; experimental study; improved; novel; particle; receptor; receptor binding; small molecule

Title: Roles of Macropinocytosis in HIV-1 infection of CD4+ T Cells Project Summary: Host cell entry is the first step in the replication cycle of HIV-1, the causative agent of AIDS. Although receptor binding of viral Env glycoprotein and subsequent virus-cell fusion during the virus entry process have been described in detail, subcellular locations of these events remain to be determined. In particular, despite the importance of CD4+ T lymphocytes as a natural host cell type for HIV-1, the contribution of virion endocytosis to HIV-1 infection of CD4+ T cells remains poorly understood. Previous studies of HIV-1 entry into T cells have largely focused on clathrin-mediated endocytosis and cell surface fusion. However, our team recently discovered that primary CD4+ T cells engage in robust macropinocytosis, a large-scale, clathrin-independent form of endocytosis. Importantly, our preliminary data indicate that pharmacological inhibition of macropinocytosis blocks both internalization of HIV-1 particles and productive infection in primary human CD4+ T cells. In contrast, the same treatment does not affect HIV infection of a commonly used CD4+ T cell line. These results suggest that the macropinocytic entry is a hitherto overlooked mode of HIV-1 entry that contributes to infection of primary human CD4+ T cells. Interestingly, our preliminary study also showed that inhibition of macropinocytosis after completion of virus internalization also blocks expression of virus-encoded genes, suggesting that macropinocytosis has another later role in productive HIV infection, which is separable from virus internalization. In the current application, we propose to determine the roles of macropinocytosis in productive infection of primary CD4+ T cells. Our long-term goal is to fully describe cellular mechanisms that promote HIV-1 infection and to use the obtained knowledge for developing antiviral strategies. Our central hypothesis in this application is that macropinocytosis in primary human CD4+ T cells contributes to productive infection as a major HIV-1 entry pathway and as a cellular function necessary for maintaining a permissive environment for the virus. To test this hypothesis, we plan to determine: 1) the extent to which macropinocytic internalization contributes to productive HIV entry into primary human CD4+ T cells; 2) the roles played by Env-receptor interactions in promoting HIV-1 macropinocytosis; 3) the influence of macropinosome environment on HIV-1 entry; and 4) the nature of post-internalization macropinocytosis function that promotes productive infection. The outcomes of the experiments outlined in this proposal will likely fill the knowledge gap in our understanding of HIV-1 entry, a fundamental aspect of the HIV-1 replication cycle, and help us develop or improve antiviral strategies that target virus entry and establishment of productive infection in CD4+ T cells.

标题：大型细胞增多症在CD4+ T细胞HIV-1感染中的作用 项目摘要： 宿主细胞进入是HIV-1复制周期的第一步，HIV-1是AIDS的致病药物。虽然受体 病毒ENV糖蛋白的结合和随后在病毒进入过程中的病毒 - 细胞融合一直是 详细描述，这些事件的亚细胞位置仍有待确定。特别是，尽管有 CD4+ T淋巴细胞作为HIV-1的天然宿主细胞类型的重要性，病毒体内吞作用的贡献 CD4+ T细胞的HIV-1感染仍然很少了解。先前对HIV-1进入T细胞的研究已有 主要集中于网格蛋白介导的内吞作用和细胞表面融合。但是，我们的团队最近 发现主要的CD4+ T细胞从事强大的大型细胞增多症，这是一种大规模的，非clathrin独立的 内吞作用的形式。重要的是，我们的初步数据表明药理抑制 大型细胞增多症阻止了原发性人的HIV-1颗粒的内在化和生产性感染 CD4+ T细胞。相比之下，相同的治疗不会影响常用的CD4+ T细胞的HIV感染 线。这些结果表明，大型细胞入口是迄今一种被忽略的HIV-1进入模式 导致原代人CD4+ T细胞的感染。有趣的是，我们的初步研究也表明 抑制病毒内在化完成后大型细胞增多症也阻塞了病毒编码的表达 基因，表明大型细胞增多症在生产性HIV感染中具有后来的作用，这是可分离的 来自病毒内在化。在当前的应用中，我们建议确定大型细胞增多症的作用 初级CD4+ T细胞的生产性感染。我们的长期目标是完全描述细胞机制 促进HIV-1感染并利用获得的知识来制定抗病毒药策略。我们的中心 该应用中的假设是原代人CD4+ T细胞中的巨细胞增多症有助于生产性 感染是主要的HIV-1进入途径，是维持允许性所必需的细胞功能 病毒的环境。为了检验这一假设，我们计划确定：1）大型细胞的程度 内部化有助于生产性HIV进入原代人CD4+ T细胞。 2）扮演的角色 Env受体相互作用在促进HIV-1大型细胞增多症中； 3）大型刺体的影响 HIV-1进入环境； 4）促进内部内部化后大细胞增多症功能的性质 生产感染。该提案中概述的实验结果可能会填补知识差距 在我们对HIV-1进入的理解，这是HIV-1复制周期的基本方面，并帮助我们发展 或改善针对CD4+ T细胞中生产感染的病毒进入和建立生产性感染的抗病毒策略。